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Protease-producing bacteria isolated from sub-Antarctic marine sediments of Isla de Los Estados (Argentina) were characterized, and the thermal inactivation kinetics of their extracellular proteases compared. Isolates were affiliated with the genera Pseudoalteromonas, Shewanella, Colwellia, Planococcus, and a strain to the family Flavobacteriaceae. Colwellia strains were moderate psychrophiles (optimal growth at about 15 degrees C, maximum growth temperature at around 25 degrees C). 16S rRNA phylogenetic analysis revealed that these strains and Colwellia aestuarii form a distinct lineage within the genus. The remaining isolates were psychrotolerant and grew optimally between 20 and 25 degrees C; two of them represent potentially novel species or genus (16S rRNA < 97% sequence similarity). The thermostability of the extracellular proteases produced by the isolates was analysed, and the inactivation rate constant (k (in)), the activation energy (Ea(in)) and the activation Gibbs free energy of thermal inactivation (Delta G( * ) (in)) determined. Delta G( * ) (in), calculated at 30 degrees C, varied between 97 and 124 kJ/mol. Colwellia enzyme extracts presented the highest thermosensitivity, while the most thermostable protease activity was shown by Shewanella spp. These results demonstrated that the stability to temperature of these enzymes varies considerably among the isolates, suggesting important variations in the thermal properties of the proteases that can coexist in this environment.

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Chitosan is a very useful marine polysaccharide that forms structural components in the exoskeleton of crustaceans. In this chapter, the production of chitosan (CH) and chitosan reticulated micro/nanoparticles (CHM) is described. Three case studies corresponding to different effective applications of chitosan are discussed: (i) the performance of CH to destabilize oil/water emulsion waste for water clarification. It was observed that as long as colloidal charge was maintained around zero, turbidity also showed low values and water clarification was achieved. However, when the applied doses were higher than the optimum, colloidal charge and turbidity both increased, showing emulsion restabilization. Emulsions treated with the optimum chitosan doses were clarified in very short periods; (ii) CH and CHM were used as effective antibacterial agents against three different pathogenic microorganisms that are problematic for aquaculture: Vibrio alginolyticus and parahaemolyticus, and Lactococcus garvieae and the minimum bactericidal concentrations were determined; and (iii) the removal of hexavalent chromium and the comparative performance of CH versus CHM. Results showed that at pH < 2, the adsorption capacity of CHM was higher because CH is unstable. Additionally, Cr(VI) was adsorbed on CH without further reduction; in contrast, Cr(VI) adsorbed on CHM was reduced to nontoxic Cr(III).